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[骨髓间充质干细胞治疗大鼠中风]

[Treatment of stroke in rats with bone marrow mesenchymal stem cells].

作者信息

Wei Jun-ji, Zeng Li-fen, Fan Xiao-tong, Wang Yu, Ma Wen-bin, Li Gui-lin, Dou Wan-chen, Zhang Zhen-xing, Li Shi-fang, Feng Ming, Han Qin, Li Zhao-jian, Zhang Zi-heng, Kang Jun, Kong Yan-guo, Wang Ren-zhi, Zhao Chun-hua

机构信息

Department of Neurosurgery, Peking Union Medical College Hospital, Chinese Academy of Medical Science & Peking Union Medical College, Beijing 100730, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2007 Jan 16;87(3):184-9.

Abstract

OBJECTIVE

To investigate the effects of treatment of stroke in rats with bone marrow mesenchymal stem cells (BMSCs) and mechanism thereof.

METHODS

Bone marrow of a healthy volunteer was collected and the BMSCs were separated with density gradient centrifugation. The hBMSC were cultivated and harvested until the third passage. A number of adult male Sprague-Dawley rats received corresponding behavioral training before surgery and underwent transient middle cerebral arterial occlusion (MCAO) for 2 hours. Sixty of them showing the scores of 6 approximately 12 according to the modified neurological severity score system were randomly divided into 2 groups: treatment group (n = 48, injected into the cortex around the ischemic areas with hBMSCs 3x10(5)/15 microl) and control group (n = 12, injected with D-Hanks solution 15 microl 24 hours after the establishment of MCAO models. Morris water maze test, Rotarod test and adhesive-removal test were performed since the 4th day to the 32 day after transplantation once every 3 days. 1, 2, 3, and 4 weeks after the transplantation 12 rats from each group were killed randomly to take out their brains. Immunofluorescence was used to identify the migration, survival and differentiation of the hBMSC.

RESULTS

A large number of hBMSC could be seen within 2 weeks after transplantation. The number of hBMSC decreased since the 21st day after transplantation and few cells could be found at the end of 1 month after. No definite evidence supported the differentiation of neural cells derived from the hBMSCs during the whole process. Morris water maze test showed that the mean escape time 1 week after transplantation of the treatment group was (69 +/- 10) s, significantly shorter than that of the control group [(120 +/- 0) s, P < 0.05] The significant difference persisted until the 4(th) week (P > 0.05). Rotarod test with the speed of 10 r/min showed that the mean latency period 10 days after transplantation of the treatment group was (167 +/- 18) s, significantly longer than that of the control group [(37 +/- 19) s, P < 0.05]. The significant difference persisted until the experimental terminal. The adhesive-removal test showed that the mean latency period 13 days after transplantation of the treatment group was (33 +/- 8) s, significant shorter than that of the control group [(84 +/- 13) s, P < 0.05]. The significant difference persisted until the experimental terminal.

CONCLUSION

Injection of hBMSCs into brain cortex improves neurological functional recovery after stroke. The transplanted cells can migrate and survive for a certain period, but no hBMSC express proteins phenotype of neural cells.

摘要

目的

探讨骨髓间充质干细胞(BMSCs)治疗大鼠脑卒中的效果及其机制。

方法

采集健康志愿者的骨髓,采用密度梯度离心法分离BMSCs。培养并收获人BMSCs至第3代。若干成年雄性Sprague-Dawley大鼠在手术前接受相应行为训练,然后进行大脑中动脉短暂闭塞(MCAO)2小时。其中60只根据改良神经功能缺损评分系统评分为6至12分的大鼠被随机分为2组:治疗组(n = 48,于缺血区周围皮质注射3×10(5)/15微升人BMSCs)和对照组(n = 12,在MCAO模型建立后24小时注射15微升D-Hanks液)。自移植后第4天至第32天,每3天进行一次Morris水迷宫试验、转棒试验和黏贴去除试验。移植后1、2、3和4周,每组随机处死12只大鼠并取脑。采用免疫荧光法鉴定人BMSCs的迁移、存活及分化情况。

结果

移植后2周内可见大量人BMSCs。移植后第21天起人BMSCs数量减少,移植后1个月末几乎未见细胞。整个过程中无确切证据支持人BMSCs分化为神经细胞。Morris水迷宫试验显示,治疗组移植后1周的平均逃避潜伏期为(69±10)秒,显著短于对照组[(120±0)秒,P<0.05]。该显著差异持续至第4周(P>0.05)。转棒试验中转速为10转/分钟时,治疗组移植后10天的平均潜伏期为(167±18)秒,显著长于对照组[(37±19)秒,P<0.05]。该显著差异持续至实验结束。黏贴去除试验显示,治疗组移植后13天的平均潜伏期为(33±8)秒,显著短于对照组[(84±13)秒,P<0.05]。该显著差异持续至实验结束。

结论

向脑皮质注射人BMSCs可改善脑卒中后神经功能恢复。移植细胞可迁移并存活一段时间,但无人BMSCs表达神经细胞蛋白表型。

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